research square, Year (2021-3)

Title : ( A simple micropreparative gel electrophoresis technique for purification of proteins, nucleic acids, and bioconjugates )

Authors: Sayyed Hashem Sajjadi , Shang-Jung Wu , Vitalijs Zubkovs , Hossein Ahmadzadeh , Elaheh Kafshdare Goharshadi , Ardemis A. Boghossian ,

Citation: BibTeX | EndNote

Abstract

The biochemical and biomedical fields hinge on the ability to effectively separate and purify biological macromolecules. Though this need is largely addressed with a variety of chromatographic and electrophoretic purification techniques, such techniques are usually laborious, time-consuming, and often require complex and costly instalments that are inaccessible to most laboratories. In this work, we introduce a simple micro-preparative (MP) method based on polyacrylamide gel electrophoresis (PAGE) to purify biological samples containing proteins, nucleic acids, and complex bioconjugates. Using a conventional vertical slab system, we demonstrate the extraction of purified DNA, proteins, and DNA-protein bioconjugates from their respective mixtures using MP-PAGE. We apply this system to recover DNA from a ladder mixture with yields of up to 90%, compared to the 58% yield obtained using specialized commercial devices. We also demonstrate the purification of folded enhanced yellow fluorescence protein (EYFP) from crude cell extract with 90% purity, comparable to purities achieved using a two-step size exclusion and immobilized metal-ion affinity chromatography purification procedure. Finally, we demonstrate the successful isolation of an EYFP-DNA bioconjugate sample that otherwise could not be processed using the two-step chromatography procedure. MP-PAGE thus offers a rapid and versatile means of purifying a variety of biomolecules without the need for specialized equipment.

Keywords

, Preparative electrophoresis; Protein; DNA; Protein, DNA conjugate; Chromatography.
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@article{paperid:1105285,
author = {سیدهاشم سجادی and شانگ جونگ وو and ویتالی زوبکوف and Ahmadzadeh, Hossein and Kafshdare Goharshadi, Elaheh and آردمیس آ. بوگوسیان},
title = {A simple micropreparative gel electrophoresis technique for purification of proteins, nucleic acids, and bioconjugates},
journal = {research square},
year = {2021},
month = {March},
issn = {2693-5015},
keywords = {Preparative electrophoresis; Protein; DNA; Protein-DNA conjugate; Chromatography.},
}

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%0 Journal Article
%T A simple micropreparative gel electrophoresis technique for purification of proteins, nucleic acids, and bioconjugates
%A سیدهاشم سجادی
%A شانگ جونگ وو
%A ویتالی زوبکوف
%A Ahmadzadeh, Hossein
%A Kafshdare Goharshadi, Elaheh
%A آردمیس آ. بوگوسیان
%J research square
%@ 2693-5015
%D 2021

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